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. 1982;1(12):1629–1633. doi: 10.1002/j.1460-2075.1982.tb01365.x

The translational map of the Autographa californica nuclear polyhedrosis virus (AcNPV) genome.

H Esche, H Lübbert, B Siegmann, W Doerfler
PMCID: PMC553261  PMID: 6327284

Abstract

We have mapped early and late viral gene products expressed in Autographa californica nuclear polyhedrosis virus ( AcNPV )-infected Spodoptera frugiperda cells by cell-free translation of virus-specific RNA which was selected by hybridization to cloned restriction endonuclease fragments of AcNPV DNA. Proteins synthesized in vitro were labeled with [35S]methionine and analyzed by SDS-polyacrylamide gel electrophoresis followed by fluorography. At least four early AcNPV -specific polypeptides were found which mapped in two regions of the genome (9-25 and 43-59 map units). These early mRNAs are also synthesized at late times in the infection cycle. Cell-free translation of restriction fragment-selected late AcNPV -specific RNA (24 h post-infection) resulted in the identification and mapping of 24 viral proteins. Curiously, the region between approximately 70 and 80 map units on the viral genome has been found silent with respect to mRNA which is translatable in a cell-free system. However, there may be RNA transcribed from this viral DNA segment.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Carstens E. B., Tjia S. T., Doerfler W. Infection of Spodoptera frugiperda cells with Autographa californica nuclear polyhedrosis virus I. Synthesis of intracellular proteins after virus infection. Virology. 1979 Dec;99(2):386–398. doi: 10.1016/0042-6822(79)90017-5. [DOI] [PubMed] [Google Scholar]
  3. Cochran M. A., Carstens E. B., Eaton B. T., Faulkner P. Molecular Cloning and Physical Mapping of Restriction Endonuclease Fragments of Autographa californica Nuclear Polyhedrosis Virus DNA. J Virol. 1982 Mar;41(3):940–946. doi: 10.1128/jvi.41.3.940-946.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Esche H., Schilling R., Doerfler W. In vitro translation of adenovirus type 12-specific mRNA isolated from infected and transformed cells. J Virol. 1979 Apr;30(1):21–31. doi: 10.1128/jvi.30.1.21-31.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Esche H., Siegmann B. Expression of early viral gene products in adenovirus type 12-infected and -transformed cells. J Gen Virol. 1982 May;60(Pt 1):99–113. doi: 10.1099/0022-1317-60-1-99. [DOI] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Lübbert H., Kruczek I., Tjia S., Doerfler W. The cloned EcoRI fragments of Autographa californica nuclear polyhedrosis virus DNA. Gene. 1981 Dec;16(1-3):343–345. doi: 10.1016/0378-1119(81)90092-5. [DOI] [PubMed] [Google Scholar]
  8. Miller L. K., Dawes K. P. Physical Map of the DNA Genome of Autographa californica Nuclear Polyhedrosis Virus. J Virol. 1979 Mar;29(3):1044–1055. doi: 10.1128/jvi.29.3.1044-1055.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  10. Ricciardi R. P., Miller J. S., Roberts B. E. Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4927–4931. doi: 10.1073/pnas.76.10.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Smith G. E., Summers M. D. Restriction Maps of Five Autographa californica MNPV Variants, Trichoplusia ni MNPV, and Galleria mellonella MNPV DNAs with Endonucleases SmaI, KpnI, BamHI, SacI, XhoI, and EcoRI. J Virol. 1979 Jun;30(3):828–838. doi: 10.1128/jvi.30.3.828-838.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Smith G. E., Vlak J. M., Summers M. D. In Vitro Translation of Autographa californica Nuclear Polyhedrosis Virus Early and Late mRNAs. J Virol. 1982 Oct;44(1):199–208. doi: 10.1128/jvi.44.1.199-208.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Tjia S. T., Carstens E. B., Doerfler W. Infection of Spodoptera frugiperda cells with Autographa californica nuclear polyhedrosis virus II. The viral DNA and the kinetics of its replication. Virology. 1979 Dec;99(2):399–409. doi: 10.1016/0042-6822(79)90018-7. [DOI] [PubMed] [Google Scholar]
  14. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro. 1977 Apr;13(4):213–217. doi: 10.1007/BF02615077. [DOI] [PubMed] [Google Scholar]
  15. Vlak J. M., Smith G. E. Orientation of the Genome of Autographa californica Nuclear Polyhedrosis Virus: a Proposal. J Virol. 1982 Mar;41(3):1118–1121. doi: 10.1128/jvi.41.3.1118-1121.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vlak J. M., Smith G. E., Summers M. D. Hybridization Selection and In Vitro Translation of Autographa californica Nuclear Polyhedrosis Virus mRNA. J Virol. 1981 Dec;40(3):762–771. doi: 10.1128/jvi.40.3.762-771.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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